旋毛虫T668基因真核表达载体的构建及其在BHK细胞中的表达  

Construction and expression in BHK cells of eukaryotic expression plasmid with T668 gene of Trichinella spiralis

作　　者：白雪[1] 吴秀萍[1] 王学林[1] 王鑫蕊[1] 于建立[1] 唐斌[1] 刘国兴[1] 冯爽[1] 陈根元[1] 刘明远[1] 

机构地区：[1]吉林大学人兽共患病研究所,吉林长春130062

出　　处：《中国兽医学报》2010年第4期469-472,共4页Chinese Journal of Veterinary Science

基　　金：国家杰出青年基金(NSFC30825033);国家科技传染病重大专项(2008ZX10004-011);国家自然科学基金(NSFC30771885)

摘　　要：利用RT-PCR技术从中国河南猪旋毛虫(国际标准虫种编号为ISS534)新生幼虫得到T668基因,并克隆入pEGFP-N1真核表达载体中构建重组质粒,该重组质粒在脂质体介导下转染BHK细胞。EGFP标签证明质粒DNA成功转染到细胞中并得以表达,Western blotting鉴定,细胞裂解液样品中有1条约77 000的条带,可被绿色荧光抗体及猪感染旋毛虫阳性血清所识别,与预计大小一致,表明成功构建T668-pEGFP-N1真核表达质粒,并在BHK细胞融合表达,表达产物具有抗原性。The T668 gene was amplified by RT-PCR from Trichinella spiralis NBL originated from pigs in Henan Province ISS534 and cloned into the eukaryotic expression plasmid pEGFP-N1.Then,the pEGFP-N1-T668 plasmid was transfected into BHK cells mediated by GenEscortTMⅡ.EGFP showed that the recombinant plasmid was expressed in BHK cells.Western-blotting results illustrated that there was a bands approximately in size in BHK221 cells lysate,it was recognized by the the anti-EGFP antibodies and the pig serum against T.spiralis,indicating that the T668 protein fusion expressed transiently in the cells and had antigenicity.

关 键 词：旋毛虫 T668基因 真核表达 免疫荧光 

分 类 号：S852.7[农业科学—基础兽医学]